Biphenyl derivatives

ABSTRACT

Biphenyl derivatives represented by the following formula (1):  
                 
 
     wherein R 1  represents a C 5-7  alkyl group, R 2  represents a substituted or unsubstituted aromatic hydrocarbon, or cycloalkyl group, R 3  represents a tetrazolyl group, —NHCOCF 3 , —NHSO 2 CF 3  or —SO 2 NHCONHR 4  in which R 4  represents a substituted or unsubstituted aromatic hydrocarbon group, and Z represents a single bond, a C 1-4  alkylene group or —SO 2 NH—, and salts thereof; and ACAT inhibitors and medicines containing them as active ingredients.  
     The compounds according to the present invention possess excellent ACAT inhibitory activities, and are useful as preventives and/or therapeutics for diseases cased by the enhancement of ACAT activity, for example, hypercholesterolemia, atherosclerosis and the like.

TECHNICAL FIELD

[0001] This invention relates to compounds possessing excellentacyl-CoA:cholesterol acyltransferase (hereinafter abbreviated as “ACAT”)inhibitory activities, and also to medicines containing the same.

BACKGROUND ART

[0002] Free cholesterol has important physiological activities as aconstituent of cell membranes and the precursor of bile acids, and alsoas a regulatory factor for the metabolism of cholesterol. Inhyperlipidemia characterized by an extraordinarily high value of serumcholesterol or the like, however, it has been considered thatatherosclerosis advances to result in an increase of the onset risk ofcoronary diseases. An increase of serum cholesterol has been, therefore,ranked as the greatest risk factor for coronary diseases.

[0003] ACAT is an enzyme which catalyzes esterification of cholesterolin cells, and physiologically, plays an important role in the control ofthe free cholesterol levels in blood and cells. It has been reportedthat the physiological role of ACAT differs depending on the tissue andthat ACAT takes part in the absorption of exogenous cholesterol in thesmall intestine, in the secretion of very-low-density lipoproteins(VLDL) in the liver, and in the accumulation of cholesterol esters inarterial walls. However, the enhanced ACAT activity leads to an onsetand advancement of hyperlipidemia and arteriosclerosis due to theincrease of serum lipids and the formation of foam cells based on theexcessive accumulation of cholesterol esters in arterial walls.

[0004] The inhibition of ACAT activity is, therefore, expected to bringabout lipid lowering effect on the basis of the suppression ofcholesterol absorption through the digestive tracts and the suppressionof VLDL secretion from the liver, and further, antiarterioscleroticeffect on the basis of the suppression of the formation of foam cells.Aiming at hyperlipidemia treatment agents and antiarterioscleroticagents, a variety of substances having ACAT inhibitory activity has beendeveloped accordingly. Under the current circumstances, however, theseconventional ACAT inhibitors have not found practical utility yet,because, in clinical trials, they have not been able to obtainsufficient effect or have induced side effects such as hepatopathy,degeneration or necrosis of the adrenal cortex, and diarrhea caused bythe suppression of fat absorption.

[0005] An object of the present invention is, therefore, to provide anew substance having ACAT inhibitory activity and a medicine containingthe same.

DISCLOSURE OF THE INVENTION

[0006] With the foregoing circumstances in view, the present inventorshave proceeded with an extensive investigation. As a result, it has beenfound that the compounds represented by the formula (1) described belowand salts thereof have excellent ACAT inhibitory activities and areuseful as preventives and/or therapeutics for diseases caused by theenhancement of ACAT activity, for example, hypercholesterolemia,atherosclerosis and the like, leading to the completion of the presentinvention.

[0007] Specifically, the present invention provides a biphenylderivative represented by the following formula (1):

[0008] wherein R¹ represents a C₅₋₇ alkyl group, R² represents asubstituted or unsubstituted aromatic hydrocarbon, or a cycloalkylgroup, R³ represents a tetrazolyl group, —NHCOCF₃, —NHSO₂CF₃ or—SO₂NHCONHR⁴ in which R⁴ represents a substituted or unsubstitutedaromatic hydrocarbon group, and Z represents a single bond, a C₁₋₄alkylene group or —SO₂NH—, or a salt thereof; and an ACAT inhibitor andmedicine containing them as an active ingredient.

[0009] The present invention also provides a medicinal compositioncomprising the biphenyl derivative or the salt thereof and apharmacologically acceptable carrier.

[0010] The present invention further provides a method for the treatmentof a disease caused by the enhancement of ACAT activity, which comprisesadministering the biphenyl derivative or the salt thereof.

[0011] The present invention still further provides use of the biphenylderivative or the salt thereof for the production of a medicine.

BEST MODES FOR CARRYING OUT THE INVENTION

[0012] In the formula (1), the C₅₋₇ alkyl group represented by R¹ can beeither linear or branched, although a linear alkyl group is preferred.Particularly preferred are n-pentyl, n-hexyl and n-heptyl from thestandpoint of ACAT inhibitory activity.

[0013] As the aromatic hydrocarbons represented by R and R⁴, C₆₋₁₀aromatic hydrocarbon groups are preferred, with phenyl and naphthylbeing more preferred and phenyl being particularly preferred. As thesubstituents which can substitute on the aromatic hydrocarbon groups, 1to 3 substituents selected from halogen atoms and C₁₋₅ alkyl groups arepreferred. Examples of the halogen atoms can include fluorine atom,chlorine atom, bromine atom and iodine atom, with fluorine atom beingparticularly preferred. The C₁₋₅ alkyl groups can be either linear orbranched, and their specific examples can include methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyland isopentyl. Among these, methyl, ethyl and isopropyl are particularlypreferred.

[0014] As the cycloalkyl group represented by R², C₃₋₈, especially C₅₋₇cycloalkyl groups are preferred. Specifically, cyclohexyl isparticularly preferred.

[0015] Preferred as R² are C₆₋₁₀ aromatic hydrocarbon which may besubstituted by 1 to 3 substituents selected from halogen atoms and C₁₋₅alkyl groups, or C₃₋₈ cycloalkyl groups, and more preferred are phenylwhich may be substituted by 1 to 3 substituents selected from halogenatoms and C₁₋₅ alkyl groups, or C₅₋₇ cycloalkyl groups. Of these,particularly preferred specific examples of R² can include di- ortrihalogenophenyl groups such as difluorophenyl and trifluorophenyl; di-or tri-C₁₋₅ alkylphenyl groups such as diisopropylphenyl andtrimethylphenyl; and cyclohexyl.

[0016] Preferred as R⁴ are C₆₋₁₀ aromatic hydrocarbon groups which maybe substituted by 1 to 3 substituents selected from halogen atoms andC₁₋₅ alkyl groups, and more preferred is a phenyl group which may besubstituted by 1 to 3 substituents selected from halogen atoms and C₁₋₅alkyl groups. Of these, particularly preferred specific examples of R⁴can include di- or trihalogenophenyl groups such as difluorophenyl andtrifluorophenyl; and di- or tri-C₁₋₅ alkylphenyl groups such asdiisopropylphenyl and trimethylphenyl.

[0017] As the tetrazolyl group represented by R³, 5-tetrazolyl isparticularly preferred. Particularly preferred examples of R³ caninclude tetrazolyl, —NHCOCF₃ and —NHSO₂CF₃.

[0018] Illustrative of the C₁₋₄ alkylene group represented by Z aremethylene, ethylene, 1-methylethylene, trimethylene, and tetramethylene,with methylene being particularly preferred. Particularly preferredexamples of Z can include a single bond, methylene and —SO₂NH—.

[0019] No particular limitation is imposed on the salt of the biphenylderivative represented by the formula (1) [the invention compound (1)],provided that it is physiologically acceptable. Preferred examples,however, can include mineral acid salts such as the hydrochloride,sulfate, phosphate and nitrate; organic acid salts such as the citrate,oxalate, fumarate, maleate, formate, acetate, methanesulfonate,benzenesulfonate and paratoluenesulfonate; the carbonate; alkali metalsalts such as the sodium and potassium salts; alkaline earth salts suchas the calcium and magnesium salts; and the ammonium salt.

[0020] In the present invention, the invention compound (1) or its saltincludes its internal salts, adducts, complexes, solvates, hydrates andthe like.

[0021] The invention compound (1) or its salt can be produced, forexample, in accordance with the following reaction scheme:

[0022] wherein R^(3a) represents a group, which can be converted intoR³, or R³ itself; Y represents —NHCOOR⁵, —NCO, —NHCOCH₂X¹ or—NHCONHSO₂X² in which R⁵ represents an alkyl or phenyl group and X¹ andX² represent halogen atoms; and R¹, R² and Z have the same meanings asdefined above.

[0023] Namely, the invention compound (1) can be obtained by reactingthe compound (3) with the secondary amine (2) to obtain the compound(1a) and then converting R^(3a) into R³ as needed.

[0024] The secondary amine (2) can be obtained, for example, bycondensing an alkylamine with a substituted biphenyl methylhalide. As analternative, it can also be obtained by reducing anN-alkanoyl-N-substituted biphenyl methylamine, which has been obtainedby a reaction between a substituted biphenyl methylamine and a fattyacid halide, with a reducing agent such as lithium aluminum hydride.

[0025] As the compound (3), on the other hand, the group described abovecan be mentioned. For a specific description, use of a carbamate(R²NHCOOR⁵) or isocyanate (R²NCO) as the compound (3) affords a compoundin which Z is a single bond, use of R²—NHCOCH₂X¹ as the compound (3)yields a compound in which Z is a methylene group, and use ofR²—NHCONHSO₂X² as the compound (3) provides a compound in which Z is—SO₂NH—.

[0026] The condensation reaction between the secondary amine (2) and thecompound (3) can be conducted, for example, in the presence of a basesuch as a tertiary amine or potassium carbonate, although the reactionconditions vary depending on the kind of Y in the compound (3).

[0027] As R^(3a), N-protected tetrazolyl groups, nitro group and thelike can be mentioned in addition to R³. As an illustrative N-protectedtetrazolyl group, N-triphenylmethyltetrazolyl can be mentioned. Thedeprotection reaction of the protecting group can be conductedpreferably by treatment with an acid such as hydrochloric acid. IfR^(3a) is a nitro group, a compound in which R³ is —NHCOCF₃ or —NHSO₂CF₃can be obtained when, after convension of the nitro group into an aminogroup by reduction, the amino group is trifluoroacetylated ortrifluoromethylsulfonylated.

[0028] It is to be noted that the production of the invention compound(1) is not limited to the reaction scheme relying upon the reactionbetween the secondary amine (2) and the compound (3). As any reactionscheme can be employed insofar as a biphenylmethyl group, R¹ and—ZCONHR² can bind together via a nitrogen atom, it is possible tointroduce the biphenylmethyl group or R¹ into the nitrogen atom at last.

[0029] The salt of the invention compound (1) can be produced, forexample, by mixing the invention compound (1) with an acid or base in apolar solvent and/or a non-polar solvent.

[0030] The invention compound (1) or the salt thereof, which has beenobtained as described above, shows excellent ACAT inhibitory activityand hence, is useful as an ACAT activity inhibitor.

[0031] As an ACAT activity inhibitor can reduce the absorption ofcholesterol from food, can suppress the secretion of VLDL from the liverand can decrease the accumulation of intracellular cholesterol esters inthe walls of blood vessels such as arteries, it can lower thecholesterol level in blood and further, can prevent the formation oflesion parts due to atherosclerosis or the like. Accordingly, theinvention compound (1) or its salt is effective for diseases caused bythe enhancement of ACAT activity, for example, hypercholesterolemia,atherosclerosis, and various diseases caused by such diseases in mammals(for example, men, mice, rats, rabbits, dogs, monkeys, and the like),and is effective as preventives and/or therapeutics for such diseases.Incidentally, specific examples of hypercholesterolemia,atherosclerosis, and various diseases caused by such diseases caninclude hyperlipidemia, arteriosclerosis, cervical or cerebralarteriosclerosis, cerebrovascular disease, cerebral infarction, stroke,reperfusion injuries, ischemic heart diseases, myocardial infarction,coronary sclerosis, nephrosclerosis, arteriosclerotic nephrosclerosis,arteriolar nephrosclerosis, malignant nephrosclerosis, ischemicenteropathy, acute mesenteric vessel occlusion, chronic intestinalangina, ischemic colitis, aortic aneurysm, arteriosclerosis obliterans(ASO), and fatty liver.

[0032] The medicine according to the present invention contains theinvention compound (1) or its salt as an active ingredient, and can beused either singly or in combination with one or more other medicinalingredients or one or more medicinal ingredients having different actionmechanisms. Further, the invention compound (1) or its salt can also beadministered as a medicinal composition with a pharmacologicallyacceptable carrier added therein as needed, and hence, can be formulatedinto a medicinal composition (preparation). Namely, the medicinalcomposition according to the present invention is characterized in thatit contains the compound represented by the formula (1) and/or its salt.Examples of the term “composition (preparation)” as used herein caninclude inhalations, injections, oral preparations, perrectalpreparations, and transdermal preparations. Pharmacologically acceptablecarriers which can be incorporated in these compositions are, forexample, excipients, binders, coating materials, lubricants,sugarcoating materials, disintegrators, extenders, correctives,emulsifiers, solubilizers, dispersants, stabilizer, pH adjusters, andisotonicities.

[0033] The preferred dosage of the medicine according to the presentinvention differs depending on the condition, the kind of disease, thesex, the age, the physique and the like, but in terms of the inventioncompound (1) or its salt, it is generally preferred to administer 1 to1,000 mg in one to several portions per adult and day.

EXAMPLES

[0034] The present invention will hereinafter be described in furtherdetail on the basis of Examples, needless to say, the present inventionshall by no means be limited only to the following Examples.

Example 1

[0035]N-(2,6-Diisopropylphenyl)-N′-pentyl-N′-[[2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]urea

[0036] n-Pentylamine (129.6 mg) and triethylamine (170 mg) weredissolved in dimethylformamide (3 mL) and under ice bath cooling,[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]bromide (399.8 mg) was added, and then the mixture was stirred at roomtemperature. Forty-eight hours later, ethyl acetate (30 mL) was added,and the resulting mixture was washed three times with brine (50 mL,each). The organic layer was dried over anhydrous sodium sulfate andconcentrated under reduced pressure. The residue and phenyl2,6-diisopropylphenylcarbamate (216.9 mg) were dissolved in toluene (10mL), and the solution was heated under reflux. Two hours later, thereaction mixture was concentrated under reduced pressure. The residuewas subjected to chromatography on a silica gel column (hexane/ethylacetate) to affordN-(2,6-diisopropylphenyl)-N′-pentyl-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]urea(88.6 mg). Yield: 16%.

[0037]¹H-NMR (CDCl₃, ppm):

[0038] 0.83-0.98(m,6H), 1.23-1.42(m,13H),

[0039] 1.62-1.70(m,2H), 2.90-3.03(m,2H),

[0040] 3.33(t,2H,J=7.29 Hz), 4.49(s,2H), 5.56(s,1H),

[0041] 6.90-6.94(m,6H), 7.10-7.17(m,6H), 7.22-7.37(m,1H),

[0042] 7.42-7.54(m,2H), 7.96(dd,1H,J=7.02 Hz,J=2.43 Hz).

[0043]N-(2,6-Diisopropylphenyl)-N′-pentyl-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]urea(330 mg) was dissolved in tetrahydrofuran (4 mL). After addition of 10%hydrochloric acid (1.2 mL), the resulting mixture was stirred at roomtemperature. Seventeen hours later, the reaction mixture wasconcentrated at low temperature under reduced pressure. Water (10 mL)was added to the concentrate, followed by extraction three times withchloroform (12 mL, each). The organic layer was washed with brine (15mL), dried over anhydrous sodium sulfate, and then concentrated underreduced pressure. The residue was subjected to chromatography on asilica gel column (chloroform/methanol) to afford the compoundrepresented by the formula (4) (0.21 g). Yield: 93%.

[0044]¹H-NMR (CDC13, ppm):

[0045] 0.91(t,3H,J=6.75 Hz), 0.97-1.12(m,12H),

[0046] 1.31-1.42(m,4H), 1.60-1.78(m,2H), 2.91-3.01(m,2H),

[0047] 3.27(t,2H,J=7.83 Hz), 4.45(s,2H), 5.71(s,1H),

[0048] 6.96-7.20(m,7H),7.39-7.60(m,3H),

[0049] 7.81(dd,1H,J=7.29 Hz,J=1.62 Hz).

[0050] m.p. 129-132° C.

[0051] IR (cm⁻¹) (KBr):

[0052] 1255, 1510, 1609, 2868, 2930, 2960.

Example 2

[0053]N′-Pentyl-N′-[[2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]-N-(2,4,6-trimethylphenyl)urea

[0054] Using n-pentylamine (304.2 mg), triethylamine (442.2 mg),[[2′-(N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]bromide (400 mg) and phenyl 2,4,6-trimethylphenylcarbamate (233.8 mg),N′-pentyl-N-(2,4,6-trimethylphenyl)-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]urea(113 mg) was obtained by a similar procedure as in Example 1. Yield:22%.

[0055]¹H-NMR (CDCl₃, ppm):

[0056] 0.88(t,3H,J=6.48 Hz), 1.18-1.35(m,4H),

[0057] 1.61-1.69(m,2H), 2.05(s,3H), 2.23(s,6H),

[0058] 3.30(t,2H,J=7.29 Hz), 4.49(s,2H), 5.54(s,1H),

[0059] 6.83-6.99(m,8H), 7.12-7.16(m,3H), 7.25-7.38(m,11H),

[0060] 7.44-7.92(m,2H),

[0061] 7.94(dd,1H,J=8.91 Hz,J=2.16 Hz).

[0062]N′-Pentyl-N-(2,4,6-trimethylphenyl)-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]urea(113 mg) was treated with hydrochloric acid in a similar manner as inExample 1 to afford the compound represented by the formula (5) (61.8mg).

[0063] Yield: 82%.

[0064]¹H-NMR (CDCl₃, ppm):

[0065] 0.88(t,3H,J=6.75 Hz), 1.22-1.35(m,4H),

[0066] 1.55-1.72(m,2H), 2.00(s,6H), 2.14(s,3H),

[0067] 3.21(t,2H,J=7.29 Hz), 4.23(s,2H), 5.66(s,1H),

[0068] 6.55(s,2H), 7.07-7.29(m,5H), 7.44-7.62(m,2H),

[0069] 7.84(dd,1H,J=7.29 Hz,J=1.35 Hz).

[0070] m.p. 106-109° C.

[0071] IR (cm⁻¹) (KBr):

[0072] 1240, 1466, 1600, 2928.

Example 3

[0073]N′-Pentyl-N′-[[2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]-N-(2,4,6-trifluorophenyl)urea

[0074] According to the procedure of Example 1, a secondary amine wasproduced from n-pentylamine (0.94 g) and[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]bromide (1.50 g). Using the secondary amine and phenyl2,4,6-trifluorophenylcarbamate (0.72 g),N′-pentyl-N-(2,4,6-triflurophenyl)-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]ureawas afforded (0.50 g). Yield: 25%.

[0075]¹H-NMR (CDCl₃, ppm):

[0076] 0.89(t,3H,J=6.48 Hz), 1.23-1.36(m,4H),

[0077] 1.50-1.66(m,2H), 3.21(t,2H,J=7.56 Hz), 4.49(s,2H),

[0078] 5.54(s,1H), 6.65-6.73(m,2H), 6.90-7.53(m,22H),

[0079] 7.94(dd,1H,J=7.02 Hz,J=1.89 Hz).

[0080]N′-Pentyl-N-(2,4,6-triflurophenyl)-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]urea(173.2 mg) was treated with hydrochloric acid in a similar manner as inExample 1 to afford the compound represented by the formula (6) (60.2mg).

[0081] Yield: 52%.

[0082] H-NMR (CDCl₃, ppm):

[0083] 0.89(t,3H,J=6.48 Hz), 1.26-1.31(m,4H),

[0084] 1.55-1.65(m,2H), 3.28(t,2H,J=7.02 Hz), 4.41(s,2H),

[0085] 6.14(s,1H), 6.51-6.57(m,2H), 6.90-7.65(m,7H),

[0086] 7.82(d,1H,J=7.29 Hz).

[0087] m.p. 103-107° C.

[0088] IR (cm⁻¹) (KBr):

[0089] 1449, 1521, 1612, 1629, 2931, 2958.

Example 4

[0090]N′-Heptyl-N′-[[2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]-N-(2,4,6-trifluorophenyl)urea

[0091] 2,4,6-Trifluorobenzoic acid (148.8 mg), diphenylphosphoryl azide(223.3 mg) and triethylamine (105.1 mg) were added to benzene (8 mL) andthe mixture was heated under reflux. Fifty minutes later, a solution ofN-heptyl-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(393.7 mg) in benzene (3 mL) was added to the reaction mixture, andunder reflux, heating was continued. Upon elapsed time of 45 minutesafter the addition of the solution, the reaction mixture wasconcentrated under reduced pressure. Ethyl acetate (40 mL) was added tothe residue, and the resulting mixture was washed successively with 5%hydrochloric acid, water, a saturated sodium bicarbonate solution andbrine (40 mL, each). The organic layer was dried over anhydrous sodiumsulfate, and then concentrated under reduced pressure. The residue wassubjected to chromatography on a silica gel column (hexane/ethylacetate) to affordN′-heptyl-N-(2,4,6-trifluorophenyl)-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]ureaas a colorless amorphous mass (369.1 mg).

[0092] Yield: 74%.

[0093]¹H-NMR (CDCl₃, ppm):

[0094] 0.87(t,3H,J=6.21 Hz), 1.15-1.35(m,8H),

[0095] 1.50-1.60(m,2H), 3.20(t,2H,J=7.29 Hz), 4.48(s,2H),

[0096] 5.62(s,1H), 6.62-6.76(m,2H), 6.89-6.98(m,6H),

[0097] 7.06-7.16(m,4H), 7.22-7.53(m,12H),

[0098] 7.93(dd,1H,J=7.02 Hz,J=1.62 Hz).

[0099]N′-Heptyl-N-(2,4,6-trifluorophenyl)-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]urea(437.5 mg) was treated with hydrochloric acid in a similar manner as inExample 1 to afford the compound represented by the formula (7) (219.1mg).

[0100] Yield: 73%.

[0101] 1H-NMR (CDCl₃, ppm):

[0102] 0.88(t,3H,J=6.48 Hz), 1.15-1.47(m,8H),

[0103] 1.55-1.75(m,2H), 3.10(t,2H,J=7.56 Hz), 4.54(s,2H),

[0104] 5.89(s,1H), 6.58(dd,2H,J=7.29 Hz,J=8.37 Hz),

[0105] 7.11-7.26(m,4H), 7.40-7.61(m,3H),

[0106] 7.93(dd,1H,J=7.29 Hz,J=1.62 Hz).

[0107] m.p. 86-89° C.

[0108] IR (cm⁻¹) (KBr):

[0109] 1521, 1615, 1625, 2929.

Example 5

[0110]N-(2,4-Difluorophenyl)-N′-heptyl-N′-[[2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]urea

[0111] n-Heptylamine (3.96 g) and potassium carbonate (0.91 g) wereadded to dimethylformamide (60 mL), and under ice cooling,[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]bromide (4.00 g) was added. The resulting mixture was stirred at thesame temperature for a while, and was then stirred at room temperature.Forty-five hours later, ethyl acetate (150 mL) was added. The mixturewas washed once with water (500 mL) and four times with brine (500 mL,each), dried over anhydrous sodium sulfate, and then concentrated underreduced pressure. Toluene (100 mL) and 2,4-difluorophenyl isocyanate(1.33 g) were added to the residue and the mixture was heated underreflux. Eighty minutes later, the mixture was diluted with toluene (100mL), and then washed successively with a saturated sodium bicarbonatesolution (100 mL) and brine (100 mL). The organic layer was dried overanhydrous sodium sulfate and then concentrated under reduced pressure.The residue was subjected to chromatography on a silica gel column(hexane/ethyl acetate) to affordN-(2,4-difluorophenyl)-N′-heptyl-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]ureaas a pale yellow oil (3.15 g). Yield: 59%.

[0112]¹H-NMR (CDCl₃, ppm):

[0113] 0.88(t,3H,J=6.48 Hz), 1.15-1.35(m,8H),

[0114] 1.57-1.60(m,2H), 3.19(t,2H,J=7.29 Hz), 4.46(s,2H),

[0115] 6.37(d,1H,J=3.24 Hz), 6.73-6.93(m,11H),

[0116] 7.04-7.53(m,13H), 7.94(dd,1H,J=7.56 Hz,J=1.62 Hz),

[0117] 8.02-8.11(m,1H).

[0118]N-(2,4-Difluorophenyl)-N′-heptyl-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]urea(1.05 g) was treated with hydrochloric acid in a similar manner as inExample 1 to afford the compound represented by the formula (8) (0.34g). Yield: 59%.

[0119]¹H-NMR (CDCl₃, ppm):

[0120] 0.88(t,3H,J=6.48 Hz), 1.22-1.33(m,8H),

[0121] 1.58-1.75(m,2H), 3.30(t,2H,J=7.83 Hz), 4.53(s,2H),

[0122] 6.45(d,1H,J=3.51 Hz), 6.70-6.87(m,2H),

[0123] 7.14-7.32(m,4H), 7.34-7.65(m,3H), 7.76-7.85(m,1H),

[0124] 7.98(d,1H,J=7.56 Hz)

[0125] IR (cm⁻¹) (KBr):

[0126] 1516, 1624, 1612.

Example 6

[0127]N-Cyclohexyl-N′-heptyl-N′-[[2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]urea

[0128] Using n-heptylamine (192 mg),[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]bromide (843 mg) and cyclohexyl isocyanate (230 mg),N-cyclohexyl-N′-heptyl-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]urea(0.53 g) was obtained in a similar manner as in Example 5. Yield: 49%.

[0129]¹H-NMR (CDCl₃, ppm):

[0130] 0.87(t,3H,J=6.75 Hz), 0.94-1.40(m,12H),

[0131] 1.48-1.62(m,6H), 1.85-1.90(m,2H),

[0132] 3.06(t,2H,J=7.83 Hz), 3.61-3.71(m,1H), 4.13(d,1H),

[0133] 4.34(s,2H), 6.86-6.94(m,6H), 7.00-7.12(m,4H),

[0134] 7.17-7.53(m,12H), 7.91(dd,1H,J=6.75 Hz,J=1.35 Hz).

[0135]N-Cyclohexyl-N′-heptyl-N′-[[2′-[N-(triphenylmethyl)tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]urea(0.53 g) was treated with hydrochloric acid in a similar manner as inExample 1 to afford the compound represented by the formula (9) (0.24g). Yield: 69%.

[0136]¹H-NMR (CDCl₃, ppm):

[0137] 0.87(t,3H,J=6.75 Hz), 0.93-1.38(m,15H),

[0138] 1.51-1.66(m,3H), 1.72-1.80(m,2H),

[0139] 3.06(t,2H,J=7.83 Hz), 3.35-3.52(m,1H),

[0140] 4.14(d,1H,J=7.29 Hz), 4.37(s,2H), 7.12(s,4H),

[0141] 7.42-7.65(m,3H), 7.90(dd,1H,J=6.75 Hz,J=1.35 Hz).

[0142] IR (cm⁻¹) (KBr):

[0143] 1532, 2930.

Example 7

[0144]N′-Pentyl-N′-[[2′-[[(trifluoromethyl)carbonyl]-amino]-1,1′-biphenyl-4-yl]methyl]-N-(2,4,6-trifluorophenyl)urea

[0145] n-Amylamine (0.54 g) and potassium carbonate (0.98 g) weresuspended in dimethylformamide (3.5 mL), and[[2′-nitro-1,1′-biphenyl-4-yl]methyl] bromide (0.51 g) was added inportions. Four hours later, ethyl acetate (20 mL) was added, and theresulting mixture was washed once with water (40 mL) and twice withbrine (90 mL, each). The organic layer was dried over anhydrous sodiumsulfate and then concentrated under reduced pressure. The residue wassubjected to chromatography on a silica gel column (chloroform/ethylacetate) to afford N-[(2′-nitro-1,1′-biphenyl-4-yl)methyl]-N-pentylamineas a yellow oil (0.40 g). Yield: 77%.

[0146] Using 2,4,6-trifluorobenzoic acid (0.43 g), diphenylphosphorylazide (0.68 g), triethylamine (0.25 g) andN-[(2′-nitro-1,1′-biphenyl-4-yl)methyl]-N-pentylamine (0.40 g),N′-[(2′-nitro-1,1′-biphenyl-4-yl)methyl]-N′-pentyl-N-(2,4,6-trifluorophenyl)urea(0.60 g) was obtained by a similar procedure as in Example 4. Yield:95%.

[0147]¹H-NMR (CDCl₃, ppm):

[0148] 0.94(t,3H,J=7.02 Hz), 1.20-1.49(m,4H),

[0149] 1.59-1.75(m,2H), 3.42(t,2H,J=7.83 Hz), 4.64(s,2H),

[0150] 5.74(s,1H), 6.64-6.87(m,4H), 7.09-7.65(m,6H).

[0151]N′-[(2′-Nitro-1,1′-biphenyl-4-yl)methyl]-N′-pentyl-N-(2,4,6-trifluorophenyl)urea(196.0 mg) and anhydrous tin(II) chloride (401.6 mg) were added toethanol (4 mL), and the mixture was heated under reflux. One hour later,the solvent was distilled off, and ethyl acetate (15 mL) and a saturatedsodium bicarbonate solution (15 mL) were added to the residue. Theresultant precipitate was filtered off through Celite. The filtrate wasextracted with ethyl acetate. The organic layer was dried over anhydroussodium sulfate and concentrated under reduced pressure to affordN′-[(2′-amino-1,1′-biphenyl-4-yl)methyl]-N′-pentyl-N-(2,4,6-trifluorophenyl)urea(179.2 mg). Yield: 94%.

[0152]N′-[(2′-Amino-1,1′-biphenyl-4-yl)methyl]-N′-pentyl-N-(2,4,6-trifluorophenyl)urea(179.2 mg) was dissolved in pyridine (1.5 mL), and after addition oftrifluoroacetic anhydride (0.15 mL), the mixture was stirred at roomtemperature. Thirty minutes later, ethyl acetate (15 mL) was added, andthe resulting mixture was washed successively with water (15 mL), brine(30 mL), a saturated sodium bicarbonate solution (30 mL) and brine (30mL). The organic layer was dried over anhydrous sodium sulfate and thenconcentrated under reduced pressure. The residue was subjected tochromatography on a silica gel column (hexane/ethyl acetate) to affordthe compound represented by the formula (10) as a colorless amorphousmass (129.5 mg). Yield: 58%.

[0153]¹H-NMR (CDCl₃, ppm):

[0154] 0.94(t,3H,J=7.02 Hz), 1.32-1.78(m,4H),

[0155] 1.62-1.72(m,2H), 3.41(t,2H,J=7.29 Hz), 4.68(s,2H),

[0156] 5.73(s,1H), 6.66-6.77(m,2H), 7.20-7.49(m,7H),

[0157] 7.95(bs,1H), 8.23(d,1H,J=80.10 Hz).

[0158] IR (cm⁻¹) (KBr):

[0159] 1202, 1521, 1636.

Example 8

[0160]N′-Pentyl-N′-[[2′-[[(trifluoromethyl)sulfonyl]-amino]-1,1′-biphenyl-4-yl]methyl]-N-(2,4,6-trifluorophenyl)urea

[0161]N′-[(2′-Amino-1,1′-biphenyl-4-yl)methyl]-N′-pentyl-N-(2,4,6-trifluorophenyl)urea(100 mg) was dissolved in dichloromethane (2 mL), and while maintainingthe solution at −10 to −5° C., a solution of trifluoromethanesulfonicacid anhydride (70 mg) in dichloromethane (2 mL) was added dropwise.After the resulting mixture was stirred at the same temperature for 4hours, chloroform (20 mL) was added and the mixture was washed with asaturated sodium bicarbonate solution (20 mL). The aqueous layer wasextracted further with chloroform (10 mL). The combined organic layerswere dried over anhydrous sodium sulfate and then concentrated underreduced pressure. The residue was purified by chromatography on a silicagel column (chloroform/methanol) to afford the compound represented bythe formula (11) (60 mg). Yield: 46%.

[0162]¹H-NMR (CDCl₃, ppm):

[0163] 0.92(t,3H,J=6.75 Hz), 1.32-1.38(m,4H),

[0164] 1.68-1.78(m,2H), 3.41(t,2H,J=7.56 Hz), 4.68(s,2H),

[0165] 5.73(s,1H), 6.66-6.77(m,2H), 7.31-7.49(m,7H),

[0166] 7.95(s,1H), 8.28(d,1H,J=7.83 Hz).

[0167] IR (cm⁻¹) (KBr):

[0168] 1202, 1521, 1636.

Example 9

[0169]N′-[[2′-[[[[(2,6-Diisopropylphenyl)amino]-carbonyl]amino]sulfonyl]-1,1′-biphenyl-4-yl]methyl]-N′-heptyl-N-(2,4,6-trifluorophenyl)urea

[0170] [[2′-[(t-Butylamino)sulfonyl]-1,1′-biphenyl-4-yl]methyl] bromide(0.94 g), n-heptylamine (0.28 g) and potassium carbonate (0.35 g) wereadded to dimethylformamide and the mixture was stirred at roomtemperature. Sixteen hours and 30 minutes later, ethyl acetate (30 mL)was added, and the resulting mixture was washed three times with brine(100 mL, each). The organic layer was dried over anhydrous sodiumsulfate and then concentrated under reduced pressure. The residue wassubjected to chromatography on a silica gel column (chloroform/ethylacetate) to affordN-[[2′-[(t-butylamino)sulfonyl]-1,1′-biphenyl-4-yl]methyl]-N-heptylamineas a yellow oil (0.43 g). Yield: 42%.

[0171]¹H-NMR (CDCl₃, ppm):

[0172] 0.89(t,3H,J=7.02 Hz), 0.99(s,9H), 1.22-1.31(m,6H),

[0173] 1.45-1.60(m,4H), 2.64(t,2H,J=6.75 Hz), 3.56(s,1H),

[0174] 4.86(s,2H), 7.30-7.65(m,7H),

[0175] 8.17(dd,1H,J=7.83 Hz,J=1.35 Hz).

[0176] Using 2,4,6-trifluorobenzoic acid (0.27 g), diphenylphosphorylazide (0.44 g), triethylamine (0.17 g) andN-[[2′-[(t-butylamino)sulfonyl]-1,1′-biphenyl-4-yl]methyl]-N-heptylamine(0.43 g),N′-[[2′-[(t-butylamino)sulfonyl]-1,1′-biphenyl-4-yl]methyl]-N′-heptyl-N-(2,4,6-trifluorophenyl)ureawas afforded as colorless crystals (0.49 g) by a similar procedure as inExample 4. Yield: 81%.

[0177]¹H-NMR (CDCl₃, ppm):

[0178] 0.89(t,3H,J=7.02 Hz), 0.95(s,9H), 1.22-1.35(m,4H),

[0179] 1.52-1.58(m,2H), 1.66-1.78(m,2H),

[0180] 3.39(t,2H,J=80.10 Hz), 3.56(s,1H), 4.66(s,2H),

[0181] 5.74(s,1H), 6.66-6.77(m,2H), 7.20-7.65(m,7H),

[0182] 8.17(dd,1H,J=7.29 Hz,J=1.08 Hz).

[0183]N′-[[2′-[(t-Butylamino)sulfonyl]-1,1′-biphenyl-4-yl]methyl]-N′-heptyl-N-(2,4,6-trifluorophenyl)urea(0.49 g) was dissolved in trifluoroacetic acid (8 mL), and afteraddition of anisole (0.36 mL), the resulting mixture was stirred at roomtemperature. Twenty-four hours later, ethyl acetate (40 mL) was added tothe reaction mixture, and the mixture was washed with a saturated sodiumbicarbonate solution (60 mL) and brine (60 mL). The organic layer wasdried over anhydrous sodium sulfate and then concentrated under reducedpressure. The residue was subjected to chromatography on a silica gelcolumn (hexane/ethyl acetate) to affordN′-[[(2′-aminosulfonyl)-1,1′-biphenyl-4-yl]methyl]-N′-heptyl-N-(2,4,6-trifluorophenyl)urea(0.40 g) as a colorless amorphous mass. Yield: 90%.

[0184]¹H-NMR (CDCl₃, ppm):

[0185] 0.89(t,3H,J=6.48 Hz), 1.23-1.35(m,8H),

[0186] 1.65-1.73(m,2H), 3.41(t,2H,J=7.29 Hz), 4.25(s,2H),

[0187] 4.66(s,2H), 5.81(s,1H), 6.66-6.72(m,2H),

[0188] 7.31-7.63(m,7H), 8.14(dd,1H,J=7.83 Hz,J=1.08 Hz).

[0189]N′-([(2′-Aminosulfonyl)-1,1′-biphenyl-4-yl]methyl]-N′-heptyl-N-(2,4,6-trifluorophenyl)urea(0.40 g) and 2,6-diisopropylphenyl isocyanate (0.22 g) were added toacetone (15 mL) and the mixture was heated under reflux. Fifty minuteslater, the reaction mixture was concentrated under reduced pressure andan aqueous solution of potassium dihydrogenphosphate was added to theresidue to adjust the pH to 4 to 5. The resulting mixture was extractedwith ethyl acetate (40 mL), and the organic layer was dried overanhydrous sodium sulfate and then concentrated under reduced pressure.The residue was subjected to chromatography on a silica gel column(hexane/ethyl acetate) to afford the compound represented by the formula(12) in the form of a colorless amorphous mass (0.41 g). Yield: 74%.

[0190]¹H-NMR (CDCl₃, ppm):

[0191] 0.88(t,3H,J=6.75 Hz), 0.99(d,12H,J=6.75 Hz),

[0192] 1.23-1.34(m,8H), 1.62-1.78(m,2H), 2.51(m,2H),

[0193] 3.43(t,2H,J=7.56 Hz), 4.67(s,2H), 5.76(s,1H),

[0194] 6.63-6.74(m,2H), 7.07(d,2H,J=80.10 Hz),

[0195] 7.16-7.66(m,8H), 8.25(d,1H,J=7.02 Hz).

[0196] m.p. 89-93° C.

[0197] IR (cm⁻¹) (KBr):

[0198] 1121, 1502, 1521, 1636.

Example 10

[0199]N-[[[(2,6-Diisopropylphenyl)amino]carbonyl]methyl]-N-pentyl-N-[[2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]amine

[0200][[2′-[N-(Triphenylmethyl)tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]bromide (2.32 g) and sodium azide (0.59 g) were suspended in a mixedsolution of dimethylformamide (8 mL) and water (0.4 mL), and then themixture was stirred at room temperature. Twenty-two hours later, ethylacetate (100 mL) was added, and the resulting mixture was washed threetimes with brine (300 mL, each). The organic layer was dried overanhydrous sodium sulfate and concentrated under reduced pressure. Theresidue was dissolved in tetrahydrofuran (40 mL), and under ice bathcooling, lithium aluminum hydride (322.0 mg) was added. The mixture wasstirred at room temperature for 1 hour, and under ice bath cooling,water (8 mL), a 10% aqueous solution of sodium hydroxide (8 mL) andwater (8 mL) were added successively. The resulting precipitate wasfiltered off, and the filtrate was concentrated under reduced pressure.Water (80 mL) was added to the residue, and the mixture was extractedtwice with chloroform (60 mL, each). The organic layers were combinedand then washed with brine (100 mL). The organic layer was dried overanhydrous sodium sulfate and concentrated under reduced pressure. Theresidue was dissolved in dichloromethane (30 mL), and after addition oftriethylamine (1500 mg) and valeroyl chloride (1.80 g), the mixture wasstirred under ice bath cooling. Forty minutes later, chloroform (50 mL)was added to the reaction mixture, and the resulting mixture was washedwith a saturated sodium bicarbonate solution (100 mL) and brine (100mL). The organic layer was dried over anhydrous sodium sulfate andconcentrated under reduced pressure. The residue was subjected tochromatography on a silica gel column (hexane/ethyl acetate) to affordN-pentanoyl-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amineas colorless crystals (1.61 g).

[0201] Yield: 67%.

[0202]¹H-NMR (CDCl₃, ppm):

[0203] 0.90(t,3H,J=7.29 Hz), 1.23-1.37(m,2H),

[0204] 1.55-1.66(m,2H), 2.10(t,2H,J=8.10 Hz),

[0205] 4.32(d,2H,J=5.40 Hz), 5.41(bs,1H), 6.88-7.53(m,22H),

[0206] 7.96(dd,1H,J=6.75 Hz,J=2.16 Hz).

[0207]N-Pentanoyl-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(1.61 g) was dissolved in tetrahydrofuran (40 mL) and at roomtemperature, lithium aluminum hydride (340.1 mg) was added, and then themixture was heated under reflux. Ninety minutes later, water (5 mL), a10% aqueous solution of sodium hydroxide (5 mL) and water (5 mL) wereadded successively under ice bath cooling. The organic layer wasseparated, dried over anhydrous sodium sulfate, and then concentratedunder reduced pressure. As water remained to the residue, ethyl acetatewas added to conduct extraction twice from the remaining water. Theorganic layer was dried over anhydrous sodium sulfate, and thenconcentrated under reduced pressure to affordN-pentyl-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine (1.57 g). Yield: 100%.

[0208] N-(Chloroacetyl)-2,6-diisopropylaniline (196.3 mg),N-pentyl-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(430.0 mg), potassium iodide (70 mg) and triethylamine (0.6 g) weresuspended in dimethylformamide (3 mL). The resulting mixture was heatedto about 80° C., and then stirred at that temperature. Three hourslater, the reaction mixture was cooled, ethyl acetate (30 mL) was added,and the mixture was washed four times with brine (50 mL, each). Theorganic layer was dried over anhydrous sodium sulfate and concentratedunder reduced pressure. The residue was subjected to chromatography on asilica gel column (hexane/ethyl acetate) to affordN-[[[(2,6-diisopropylphenyl)amino]carbonyl]methyl]-N-pentyl-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(196.4 mg). Yield: 33%.

[0209]¹H-NMR (CDCl₃, ppm):

[0210] 0.90(t,3H,J=7.02 Hz), 1.13(d,12H,J=7.02 Hz),

[0211] 1.24-1.41(m,4H), 1.55-1.65(m,2H),

[0212] 2.60(t,2H,J=7.83 Hz), 2.63-2.98(m,2H), 3.24(s,2H),

[0213] 3.62(s,2H), 6.90-6.94(m,6H), 7.07-7.55(m,19H),

[0214] 7.93(dd,1H,J=6.48 Hz,J=1.08 Hz), 8.63(s,1H).

[0215]N-[[[(2,6-Diisopropylphenyl)amino]carbonyl]methyl]-N-pentyl-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(278.3 mg) was dissolved in tetrahydrofuran (10 mL), and after additionof 10% hydrochloric acid (3.0 mL), the resulting mixture was stirred atroom temperature. Nineteen hours later, the solvent was distilled off,and to the residue, water (30 mL) was added. The mixture so obtained wasextracted twice with chloroform (30 mL, each). The organic layer wasdried over anhydrous sodium sulfate and concentrated under reducedpressure. The residue was subjected to chromatography on a silica gelcolumn (chloroform/methanol) to affordN-[[[(2,6-diisopropylphenyl)amino]carbonyl]methyl]-N-pentyl-N-[[2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]aminehydrochloride as pale yellow crystals (190.6 mg). Yield: 93%.

[0216]¹H-NMR (CDCl₃, ppm):

[0217] 0.90(t,3H,J=6.48 Hz), 1.12(d,12H,J=7.02 Hz),

[0218] 1.25-1.43(m,4H), 1.85(bs,2H), 2.89-2.99(m,2H),

[0219] 3.20(bs,2H), 3.83(bs,2H), 4.32(bs,2H),

[0220] 7.10-7.16(m,4H), 7.25-7.31(m,1H), 7.39-7.59(m,5H),

[0221] 7.91(dd,1H,J=7.29 Hz,J=1.35 Hz), 9.23(bs,1H).

[0222] m.p. 102-107° C.

[0223] IR (cm⁻¹) (KBr):

[0224] 1459, 1471, 1687, 2869, 2930, 2962.

[0225]N-[[[(2,6-Diisopropylphenyl)amino].carbonyl]methyl]-N-pentyl-N-[[2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]aminehydrochloride (140.4 mg) was suspended in a saturated sodium bicarbonatesolution (5 mL) and the mixture was stirred for 1 hour. The mixture wasadjusted to pH 3 with dilute hydrochloric acid, and then extracted withethyl acetate (30 mL). The organic layer was washed with brine. Theorganic layer was dried over anhydrous sodium sulfate and concentratedunder reduced pressure to afford the compound represented by the formula(13) as a colorless solid (107.3 mg). Yield: 82%.

[0226]¹H-NMR (CDCl₃, ppm):

[0227] 0.90(t,3H,J=6.75 Hz), 1.11(d,12H,J=7.02 Hz),

[0228] 1.23-1.34(m,4H), 2.74(t,2H,J=7.29 Hz),

[0229] 2.86-2.96(m,2H), 3.33(s,2H), 3.85(s,2H),

[0230] 7.07-7.16(m,4H), 7.25-7.31(m,3H), 7.38-7.41(m,1H),

[0231] 7.47-7.60(m,2H), 7.99(dd,1H,J=7.29 Hz,J=1.35 Hz),

[0232] 8.77(s,1H).

[0233] m.p. 74-80° C.

[0234] IR (cm⁻¹) (KBr):

[0235] 1459, 1505, 1658, 2869, 2930.

Example 11

[0236]N-Pentyl-N-[[2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]-N-[[[(2,4,6-trimethylphenyl)amino]-carbonyl]methyl]amine

[0237] N-(Chloroacetyl)-2,4,6-trimethylaniline (443.2 mg),N-pentyl-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(780.0 mg), potassium iodide (129.1 mg) and triethylamine (1.15 g) weresuspended in dimethylformamide (6 mL). The resulting mixture was heatedto about 80° C., and then stirred at that temperature. Three hourslater, the reaction mixture was cooled, ethyl acetate (50 mL) was added,and the mixture was washed four times with brine (100 mL, each). Theorganic layer was dried over anhydrous sodium sulfate and concentratedunder reduced pressure. The residue was subjected to chromatography on asilica gel column (hexane/ethyl acetate) to affordN-pentyl-N-([[(2,4,6-trimethylphenyl)amino]carbonyl]methyl]-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(395.4 mg). Yield: 39%.

[0238]¹H-NMR (CDCl₃, ppm):

[0239] 0.89(t,3H,J=7.02 Hz), 1.23-1.31(m,4H),

[0240] 1.45-1.65(m,2H), 2.08(s,6H), 2.26(s,3H),

[0241] 2.59(t,2H,J=7.83 Hz), 3.24(s,2H), 3.62(s,2H),

[0242] 6.89-7.54(m,24H), 7.92(dd,1H,J=7.02 Hz,J=1.89 Hz),

[0243] 8.59(s,1H).

[0244]N-Pentyl-N-[[[(2,4,6-ttimethylphenyl)amino]-carbonyl]methyl]-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(395.4 mg) was dissolved in tetrahydrofuran (25 mL), and after additionof 10% hydrochloric acid, the resulting mixture was stirred for 18 hoursand 30 minutes. The solvent was then distilled off under reducedpressure. To the residue, water (45 mL) was added, and the mixture wasextracted twice with ethyl acetate (45 mL, each). The organic layer wasdried over anhydrous sodium sulfate and concentrated under reducedpressure. The residue was subjected to chromatography on a silica gelcolumn (chloroform/methanol). Relevant fractions (the hydrochloride)were concentrated, and after addition of a small amount of methanol tothe residue, the mixture was stirred together with a saturated sodiumbicarbonate solution (6 mL) for 1 hour. Under ice bath cooling, thesolution was adjusted to pH 3 with dilute hydrochloric acid. Ethylacetate (30 mL) was added, and the resulting mixture was washed sixtimes with brine (50 mL, each). The organic layer was dried overanhydrous sodium sulfate and concentrated under reduced pressure. Theresidue was subjected to chromatography on a silica gel column(chloroform/methanol) to afford the compound represented by the formula(14) as colorless crystals (188.4 mg).

[0245] Yield: 69%.

[0246]¹H-NMR (CDCl₃, ppm):

[0247] 0.89(t,3H,J=6.75 Hz), 1.23-1.43(m,4H),

[0248] 1.56-1.72(m,2H), 2.02(s,6H), 2.22(s,3H),

[0249] 2.69(t,2H,J=7.29 Hz), 3.24(s,2H), 3.78(s,2H),

[0250] 6.81(s,2H), 7.04(d,2H,J=7.83 Hz),

[0251] 7.22-7.57(m,5H), 7.95(d,1H,J=7.29 Hz), 8.68(s,1H).

[0252] m.p. 73-77° C.

[0253] IR (cm⁻¹) (KBr):

[0254] 1505, 1659, 2928, 2956.

Example 12

[0255]N-Pentyl-N-[(2′-(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]-N-([[(2,4,6-trifluorophenyl)amino]-carbonyl]methyl]amine

[0256] N-(Chloroacetyl)-2,4,6-trifluoroaniline (313.0 mg),N-pentyl-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(780.0 mg), potassium iodide (230 mg) and triethylamine (2.1 g) weresuspended in dimethylformamide (12 mL). The resulting mixture was heatedto about 80° C., and then stirred at that temperature. Three hourslater, the reaction mixture was cooled, ethyl acetate (100 mL) wasadded, and the mixture was washed four times with brine (200 mL, each).The organic layer was dried over anhydrous sodium sulfate andconcentrated under reduced pressure. The residue was subjected tochromatography on a silica gel column (hexane/ethyl acetate) to affordN-pentyl-N-[[[(2,4,6-trifluorophenyl)amino]carbonyl]methyl]-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(434.8 mg). Yield: 42%.

[0257]¹H-NMR (CDCl₃, ppm):

[0258] 0.88(t,3H,J=7.02 Hz), 1.18-1.38(m,4H),

[0259] 1.48-1.60(m,2H), 2.54(t,2H,J=7.56 Hz), 3.23(s,2H),

[0260] 3.61(s,2H), 6.71-6.77(m,2H), 6.89-6.92(m,6H),

[0261] 7.05-7.13(m,4H), 7.21-7.51(m,11H),

[0262] 7.92(d,1H,J=5.94 Hz), 7.93(d,1H,J=5.94 Hz),

[0263] 8.63(s,1H).

[0264]N-Pentyl-N-[[[(2,4,6-trifluorophenyl)amino]-carbonyl]methyl]-N-[[2′-[N-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(425.8 mg) was dissolved in tetrahydrofuran (25 mL), and after additionof 10% hydrochloric acid, the resulting mixture was stirred for 18 hoursand 30 minutes. The solvent was then distilled off under reducedpressure. To the residue, water (45 mL) was added, and the mixture wasextracted twice with ethyl acetate (45 mL, each). The organic layer wasdried over anhydrous sodium sulfate and concentrated under reducedpressure. The residue was subjected to chromatography on a silica gelcolumn (chloroform/methanol). Relevant fractions (the hydrochloride)were concentrated, and after addition of a small amount of methanol tothe residue, the mixture was stirred together with a saturated sodiumbicarbonate solution (6 mL) for 1 hour. Under ice bath cooling, thesolution was then adjusted to pH 3 with dilute hydrochloric acid. Ethylacetate (30 mL) was added, and the resulting mixture was washed sixtimes with brine (50 mL, each). The organic layer was dried overanhydrous sodium sulfate and concentrated under reduced pressure. Theresidue was subjected to chromatography on a silica gel column(chloroform/methanol) to afford the compound represented by the formula(15) (185.3 mg). Yield: 63%.

[0265]¹H-NMR (CDCl₃, ppm):

[0266] 0.88(t,3H,J=6.48 Hz), 1.15-1.50(m,4H),

[0267] 1.51-1.75(m,2H), 2.73(t,2H,J=7.83 Hz), 3.31(s,2H),

[0268] 3.80(s,2H), 6.65-6.74(m,2H), 7.04(d,2H,J=7.83 Hz),

[0269] 7.21-7.27(m,2H), 7.38-7.57(m,3H),

[0270] 7.90(dd,1H,J=7.83 Hz,J=1.08 Hz), 8.93(s,1H).

[0271] m.p. 69-73° C.

[0272] IR (cm⁻¹) (KBr):

[0273] 1449, 1521, 1610, 2932, 2957.

Example 13

[0274]N-[[[[(2,6-Diisopropylphenyl)amino]carbonyl]-amino]sulfonyl]-N-pentyl-N-[2′-[(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methyl]amine

[0275] Chlorosulfonyl isocyanate (2.43 g) was dissolved in diethyl ether(10 mL), and at −15° C., the solution was added dropwise to a solutionof 2,6-diisopropylaniline in diethyl ether (15 mL). After the mixturewas stirred at the same temperature for 1 hour and 30 minutes, theresulting precipitates were collected by filtration and then washed withhexane. Colorless crystals so obtained were dried to afford[[[(2,6-diisopropylphenyl)amino]carbonyl]amino]sulfonyl chloride (4.11g). Yield: 75%.

[0276]¹H-NMR (CDCl₃, ppm):

[0277] 1.22(s,6H), 1.25(s,6H), 3.13(m,2H),

[0278] 7.20-7.39(in,3H), 7.76(s,1H)

[0279] m.p. 131-133° C.

[0280][[2′-[N-(Triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]bromide (1.08 g), n-pentylamine (0.18 g) and potassium carbonate (0.45g) were added to N,N-dimethylformamide (7 mL) and the mixture wasstirred at room temperature. Sixteen hours later, ethyl acetate (40 mL)was added. The organic layer was washed with brine (140 mL×3 times). Theorganic layer was dried over anhydrous sodium sulfate and thenconcentrated under reduced pressure. To the residue, triethylamine (0.20g) and tetrahydrofuran (11 mL) were added. Into the mixture, a solutionof [[[(2,6-diisopropylphenyl)amino]carbonyl]amino]sulfonyl chloride(4.11 g) in tetrahydrofuran (9 mL) was added dropwise at roomtemperature. Two hours later, ethyl acetate (80 mL) was added, and theresulting mixture was washed with water (80 mL) and brine (40 mL). Theorganic layer was dried over magnesium sulfate and then concentratedunder reduced pressure. The residue was subjected to chromatography on asilica gel column (hexane/ethyl acetate) to affordN-[[[[(2,6-diisopropyl-phenyl)amino]carbonyl]amino]sulfonyl]-N-pentyl-N-[[[2′-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amineas a colorless amorphous mass (0.32 g).

[0281] Yield: 20%.

[0282]¹H-NMR (CDCl₃, ppm):

[0283] 0.83(t,3H,J=6.5 Hz), 1.09-1.29(m,18H),

[0284] 3.05-3.29(m,4H), 4.39(s,1H), 4.52(s,1H),

[0285] 6.87-6.99(m,6H), 7.15-7.51(m,20H),

[0286] 7.92(dd,1H,J=2.2,5.4 Hz)

[0287]N-[[[[(2,6-Diisopropylphenyl)amino]carbonyl]amino]-sulfonyl]-N-pentyl-N-[[[2′-(triphenylmethyl)tetrazol-5-yl]-1,1′-biphenyl-4-yl]methyl]amine(0.32 g) was dissolved in tetrahydrofuran (11 mL). At room temperature,10% hydrochloric acid was added, and the resulting mixture was stirred.Seventeen hours later, ethyl acetate (60 mL) was added, and the mixturewas washed with water (60 mL) and brine (60 mL). The organic layer wasdried over anhydrous sodium sulfate and then concentrated under reducedpressure. The residue was subjected to chromatography on a silica gelcolumn (chloroform/methanol) and further, purified by preparativethin-layer chromatography (chloroform/methanol). The thus-obtainedcrystals were suspended in diisopropyl ether. Crystals were collected byfiltration, suspended in diethyl ether, and then collected again byfiltration to afford the compound represented by the formula (16)(0.5g).

[0288]¹H-NMR (CDCl₃, ppm):

[0289] 0.97(t,3H,J=6.8 Hz), 1.20(s,6H), 1.23(s,6H),

[0290] 1.40-1.42(m,4H), 1.76-1.83(m,2H), 3.00-3.16(m,1H),

[0291] 3.52(t,2H,J=7.0 Hz), 4.52(s,2H), 4.61(s,1H),

[0292] 7.13-7.31(m,5H), 7.39-7.65(m,6H).

[0293] IR (cm⁻¹) (KBr):

[0294] 1517, 1521, 2962.

[0295] Test (Evaluation of ACAT Inhibitory Activity)

[0296] (1) Preparation of the Enzyme (ACAT)

[0297] (1-1) Each rat was reared for about 3 weeks on high-cholesterolfeed (solid feed prepared by adding 1% of cholesterol, 0.3% of sodiumcholate, 0.1% of propylthiouracil and 3% of lard to a conventional feedMF).

[0298] (1-2) The liver of the rat reared in (1-1) was collected andshredded. 10 mM HEPES buffer (pH 7.4) which contained 0.25 M of sucroseand 1 mM of EDTA was added in an amount about three times as much as theweight of the liver to suspend the latter. The liver was thenhomogenized by a glass-Teflon homogenizer.

[0299] (1-3) The homogenized liver was centrifuged for 15 minutes under22000×g, and the supernatant was collected.

[0300] (1-4) The supernatant was centrifuged further for 60 minutesunder 100000×g. To the precipitates, 10 mM HEPES buffer (pH 7.4)containing 0.25 M of sucrose and 1 mM of EDTA therein was added in abouta half of the volume used in (1-2), and the precipitates were suspendedagain.

[0301] (1-5) The suspension was again centrifuged for 60 minutes under100000×g, and the resulting precipitates were collected. 10 mM HEPESbuffer (pH 7.4) containing 0.25 M of sucrose and 2 mM of DTT therein wasadded to suspend the precipitates. The suspension was stored at −80° C.

[0302] (2) Preparation of Reagents

[0303] (2-1) Reaction Buffer

[0304] 0.75 M Phosphate buffer (pH 7.4), 800 μM BSA and 100 mM DTT weremixed together in amounts of 1.0 mL, 0.5 mL and 0.1 mL, respectively,and ultrapure water (3.4 mL) was added.

[0305] (2-2) ACAT

[0306] A refrigerated sample of the enzyme was diluted with the“reaction buffer” described above to 2.5 mg protein/mL.

[0307] (2-3) Test Samples

[0308] 10⁻³ M Solutions of the compounds to be evaluated were preparedwith methanol. 3×10⁻⁴ M Solutions were each prepared by adding 50%methanol (700 μL) to the corresponding 10⁻³ M solution (300 μL). 10⁻⁴ MSolutions were each prepared by adding 50% methanol (600 μL) to thecorresponding 3×10⁻³ M solution (300 μL). Up to 10⁻⁷ M solutions (serial3-fold dilutions), the solutions were prepared by a similar procedure.

[0309] (3) Assay Method of ACAT Activity

[0310] (3-1) ACAT (20 μL), the reaction buffer (20 μL) and desired one(5 μL) of test samples were placed in a 1.5 mL test tube and incubatedat 30° C. for 10 minutes (the resulting solution will hereinafter becalled “I”).

[0311] (3-2) Reaction substrate [¹⁴C]-oleoyl CoA (5 μL) was added into“I”, and was allowed to react at 30° C. for 4 minutes.

[0312] (3-3) Four minutes later, methanol (250 μL) was added toterminate the reaction, and then, a lipid mixture (40 μL),recovery-percentage-correcting [³H]-cholesteryl oleate (10 μL) andhexane (700 μL) were added into the test tube (this solution willhereinafter be called “II”).

[0313] (3-4) “II” was stirred in a mixer, and the hexane layer (500 μL)was collected and transferred into another test tube (this solution willhereinafter be called “III”).

[0314] (3-5) “III” was evaporated to dryness, dissolved in chloroform(10 μL), and then spotted on a TLC plate. At that time, cholesteryloleate was also spotted.

[0315] (3-6) After the spots were dried, they were developed with a85:15:0.5 mixture of hexane, diethyl ether and acetic acid as adeveloper solvent, and then stained with iodine. A plate sectioncorresponding to the spot of cholesteryl oleate was cut out and placedin a vial. At the same time, the reaction substrate [¹⁴C]-oleoyl CoA (5μL) and the recovery-percentage-correcting [³H]-cholesteryl oleate (10μL) were spotted on the TLC plate, and similarly, plate sections werecut out and placed in vials, respectively.

[0316] (3-7) Subsequently, “Aquazole II” (about 10 mL) was added to thevials. After the vials were allowed to stand for a while, they weremeasured for [¹⁴C] and [³H] radioactivities.

[0317] Using the radioactivities, the % recovery of [¹⁴C]-cholesteryloleate formed by the enzymatic reaction was calculated from theradioactivity of [³H], and from the radioactivity of [¹⁴C], the yield ofcholesteryl oleate was calculated. From the results, aconcentration-response curve was prepared, and using the nonlinear leastsquare method, a pIC₅₀ value was calculated and was used as an index forACAT inhibitory activity.

[0318] The pIC₅₀ values so calculated are shown in Table 1. TABLE 1Compound PIC₅₀ Compound of the formula (4) 5.72 Compound of the formula(5) 5.65 Compound of the formula (6) 5.52 Compound of the formula (7)6.23 Compound of the formula (8) 5.49 Compound of the formula (9) 5.18Compound of the formula (10) 6.57 Compound of the formula (11) 5.71Compound of the formula (12) 6.40 Compound of the formula (13) 5.39Compound of the formula (14) 5.23 Compound of the formula (15) 5.21Compound of the formula (16) 5.51

[0319] It is understood from the above-described pIC₅₀ values that thecompounds according to the present invention possess excellent ACATinhibitory activities.

INDUSTRIAL APPLICABILITY

[0320] The invention compounds (1) have excellent ACAT inhibitoryactivities and are useful as preventives and/or therapeutics fordiseases caused by the enhancement of ACAT activity, for example,hypercholesterolemia, atherosclerosis and the like.

1. A biphenyl derivative represented by the following formula (1):

wherein R¹ represents a C₅₋₇ alkyl group, R² represents a substituted orunsubstituted aromatic hydrocarbon, or a cycloalkyl group, R³ representsa tetrazolyl group, —NHCOCF₃, —NHSO₂CF₃ or —SO₂NHCONHR⁴ in which R⁴represents a substituted or unsubstituted aromatic hydrocarbon group,and Z represents a single bond, a C₁₋₄ alkylene group or —SO₂NH—; or asalt thereof:
 2. A biphenyl derivative or a salt thereof as defined inclaim 1, wherein R² represents a C₆₋₁₀ aromatic hydrocarbon which may besubstituted by 1 to 3 substituents selected from halogen atoms and C₁₋₅alkyl groups, or C₃₋₈ cycloalkyl group, and R⁴ represents a C₆₋₁₀aromatic hydrocarbon group which may be substituted by 1 to 3substituents selected from halogen atoms and C₁₋₅ alkyl groups.
 3. Abiphenyl derivative or a salt thereof as defined in claim 1, wherein R²represents a phenyl group which may be substituted by 1 to 3substituents selected from halogen atoms and C₁₋₅ alkyl groups, or C₅₋₇cycloalkyl groups, and R⁴ represents a phenyl group which may besubstituted by 1 to 3 substituents selected from halogen atoms and C₁₋₅alkyl groups.
 4. An acyl-CoA:cholesterol acyltransferase inhibitorcomprising as an active ingredient a compound as defined in any one ofclaims 1-3.
 5. A medicine comprising as an active ingredient a compoundas defined in any one of claims 1-3.
 6. A medicine as defined in claim5, which is a preventive and/or therapeutic for a disease caused by theenhancement of acyl-CoA:cholesterol acyltransferase activity.
 7. Amedicine as defined in claim 5, which is a preventive and/or therapeuticfor hypercholesterolemia, atherosclerosis or a disease caused by thesame.
 8. A medicinal composition comprising a compound as defined in anyone of claims 1-3 and a pharmacologically acceptable carrier.
 9. Amethod for the treatment of a disease caused by the enhancement ofacyl-CoA:cholesterol acyltransferase activity, which comprisesadministering a compound as defined in any one of claims 1-3.
 10. Use ofa compound as defined in any one of claims 1-3 for the production of amedicine.